3种新型冠状病毒株灭活疫苗间S蛋白检测结果干扰情况分析

Analysis of S protein cross-interference in three different corona virus strains

目的:考察建立的HB02株、Delta株和Omicron株新型冠状病毒(以下简称新冠)S蛋白多克隆酶联免疫检测方法(ELISA法)检测3种毒株疫苗间的交叉干扰情况。方法:将新冠HB02株、Delta株、Omicron株单价疫苗配制成不同配比和不同组合的样品,分别采用HB02株、Delta株和Omicon株新冠灭活疫苗(Vero细胞)S蛋白抗原ELISA法进行检测,比较检测结果并计算干扰率,同时分析了不同毒株抗原检测试剂盒分别检测3种新冠毒株单价抗原检测结果间的干扰率。结果:由2种毒株组分按不同剂量配比的检测结果显示,当在HB02株中加入由低到高剂量比例的Delta株时,干扰率范围为4.21%~29.53%;当在HB02株中加入由低到高剂量比例的Omicron株时,干扰率范围为4.40%~12.75%;当在Delta株中加入由低到高剂量比例的HB02株时,干扰率范围为56.17%~464.21%;当在Delta株中加入由低到高剂量比例的Omicron株时,干扰率范围为6.29%~25.91%;当在Omicron株中加入由低到高剂量比例的HB02株时,干扰率范围为32.05%~337.22%;而当在Omicron株中加入由低到高剂量比例的Delta株时,干扰率范围为44.91%~252.14%。由3种毒株组分按不同剂量配比的检测结果显示,当在HB02株中加入不同剂量比例的Delta株+Omicron株时,干扰率范围为2.79%~22.86%;当在Delta株中加入不同剂量比例的HB02株+Omicron株时,干扰率范围为67.16%~267.95%;当在Omicron株中加入不同剂量比例的HB02株+Delta株时,干扰率范围为100.66%~412.54%。此外,采用HB02株抗原检测试剂盒检测单价Delta株与单价Omicron株新冠抗原的干扰率分别为70%和90%;Delta株抗原检测试剂盒检测单价HB02株与单价Omicron株新冠抗原的干扰率分别为30%和35%;Omicron株抗原检测试剂盒检测单价HB02株与Delta株新冠抗原的干扰率分别为5%和10%。结论:由于HB02株、Delta株和Omicron株新冠疫苗中的S蛋白存在很高的相似性,因此,不同毒株按不同配比混合后,使用多克隆ELISA方法进行检测时,不同组分之间存在不同程度的干扰作用。当2种毒株组分按不同剂量配比混合时,Delta株和Omicron株对HB02株检测结果都呈现正干扰,且随着Delta株剂量比增大呈逐渐递增趋势,而Omicron株的配制比增大时干扰率变化不显著;HB02株对Delta株呈现极强的正干扰,且随着HB02株剂量比增大干扰率呈逐渐递增趋势;Omicron株对Delta株呈微弱正干扰,当Omicron株的配制比增大时干扰率未见显著变化。HB02株和Delta株对Omicron株都呈现较强的正干扰,且随着HB02株或Delta株的配制比增大呈逐级递增趋势。当3种毒株组分按不同剂量配比混合后,Delta株+Omicron株对HB02株检测结果呈现正干扰,但当Delta株+Omicron株的配制比增大时干扰率未见显著变化。HB02株+Omicron株对Delta株检测结果呈现极强的正干扰,但在HB02株和Delta株配制比不变时,不论Omicron株的浓度如何变化,Omicron株对Delta株的干扰率未见明显变化,表明HB02株对Delta株呈现极强的正干扰,但Omicron株对Delta株的检测结果无明显干扰。HB02株+Delta株对Omicron株呈现较强的正干扰,且随着HB02株和Delta株的配制比增大呈逐级递增趋势。使用不同毒株新冠抗原检测试剂盒检测3种毒株单价新冠样品,其中HB02株抗原检测试剂盒对Omicron株样品干扰率最高,但是Omicron株抗原检测试剂盒能够有效检测出HB02株新冠抗原,而Delta株抗原检测试剂盒能够检测出部分HB02株和Omicron株新冠抗原。

Objective: To investigate the S protein cross-interference of the HB02 coronavirus strain, the Delta coronavirus strain and the Omicron coronavirus strain using the developed method of polyclonal enzyme-linked immunoassay(ELISA). Methods: The monovalent vaccines of HB02 strain, Delta strain and Omicron strain were prepared in different ratios and combinations. The inactivated vaccines of HB02 strain, Delta strain and Omicon strain(Vero cell) S protein antigen ELISA method was used for detection, and the detection results were compared and the interference rate was calculated. At the same time, the interference rates between the monovalent antigen detection results of the three SARS-CoV-2 strains were analyzed. Results: The results of the two strains at different doses showed that the interference rate ranged from 4.21% to 29.53% when Delta strain was added to HB02 strain along with the increasing dose. When Omicron strain was increasingly added to HB02 strian, the interference rate ranged 4.40%~12.75%, and when HB02 strain was increasingly added to Delta strain, the interference rate ranged 56.17%~464.21%. The interference rate ranged from 6.29% to 25.91% when Omicron strain was added to Delta strain from a low to high dose. When HB02 strain was added to Omicron strain, the interference rate ranged from 32.05% to 337.22%. When Delta strain was added to Omicron strain, the interference rate ranged from 44.91% to 252.14%. The results of different doses of the three strains showed that the interference rate ranged from 2.79% to 22.86% when different doses of Delta strain+Omicron strain were added to HB02 strain. When different doses of HB02+Omicron strains were added to Delta strain, the interference rate was 67.16% to 267.95%. When different doses of HB02+Delta strains were added to Omicron strain, the interference rate was 100.66%~412.54%. In addition, the interference rates of monovalent Delta strain and Omicron strain detected by HB02 antigen detection kit were 70% and 90%, respectively. The interference rates of Delta strain antigen detection kit for monovalent HB02 strain and Omicron strain were 30% and 35%, respectively. The interference rates of Omicron strain antigen detection kit for monovalent HB02 strain and Delta strain were 5% and 10% respectively. Conclusion: Due to the high similarity of S protein in the HB02, Delta and Omicron COVID-19 vaccines, different strains were mixed at series ratios, and different components had different degrees of interference. When the components of the two strains were mixed at different doses, both Delta strain and Omicron strain showed positive interference to the HB02 strain, and the Delta strain showed a gradually increasing trend with the increasing Delta strain doses, while the interference rate did not change significantly with the variation of Omicron strain dose. HB02 strain showed strong positive interference to Delta strain, and the interference rate gradually increased with the increase of HB02 strain, while Omicron strain showed weak positive interference to Delta strain, and the interference rate did not change significantly with the increase of Omicron strain dose. Both HB02 strain and Delta strain showed strong positive interference to Omicron strain, and showed a stepwise increasing trend with the increase of the composition ratio of HB02 strain or Delta strain. When the three strains were mixed at different ratios, the Delta strain+Omicron strain showed positive interference to the HB02 strain, but the interference rate did not change significantly with the increase of the composition of Delta strain+Omicron strain. HB02 strain+Omicron strain showed strong positive interference to Delta strain. However, when the composition of HB02 strain and Delta strain was unchanged, the interference of Omicron strain to Delta strain did not change significantly no matter how Omicron strain changed. The results showed that HB02 strain had strong positive interference to Delta strain, but Omicron strain did not exert obvious interference to Delta strain. HB02+Delta strain showed strong positive interference to Omicron strain, and the ratio of HB02 and Delta strain increased stepwise. Different COVID-19 antigen detection kits were used to detect the monovalent COVID-19 samples of the three virus strains. Among them, HB02 antigen detection kit had the highest interference rate to Omicron strain samples, but Omicron strain antigen detection kit could effectively detect HB02 COVID-19 antigen. The Delta strain antigen detection kit could detect some COVID-19 antigens of HB02 strain and Omicron strain.